Name: 小鱼，大问题：墨西哥 Tetra 研究的现代技术集合 (视频) | JoVE Journal
Uploaded: 2020-02-24T00:00:00.000+00:00
Duration: 2 s
Description: 了解墨西哥盲鱼（Astyanax mexicanus）研究的现代技术集合，涵盖基因操作、行为追踪、神经丘成像及整体原位杂交等方法。这些标准化方案为发育生物学、遗传学和行为神经科学研究提供了可重复的技术支持。

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5 minutes

作者感谢美国国立卫生研究院 （NH089934， DK108495） 的资助。F. Leal 和 A. Powers 的编辑感谢。

<ol><li>Mitchell, R. W., Russel, W. H., Elliot, W. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mexican+eyeless+characin+fishes%2C+Genus+Astyanax%3A+Environment%2C+Distribution%2C+and+Evolution%5BTitle%5D)+AND+%22Special+Publications%2C+the+Museum+Texas+Tech+University%22%5BJournal%5D)">Mexican eyeless characin fishes, Genus Astyanax: Environment, Distribution, and Evolution.</a> Special Publications, the Museum Texas Tech University. 12, (1977).</li>
<li>Elliott, W. R. Cave Exploration and Mapping in the Sierra de El Abra Region. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aElliott%2C+WR+%22Biology+and+Evolution+of+the+Mexican+Cavefish%22">Biology and Evolution of the Mexican Cavefish</a>. , 9-40 (2015).</li>
<li>Herman, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+role+of+gene+flow+in+rapid+and+repeated+evolution+of+cave-related+traits+in+Mexican+tetra%2C+Astyanax+mexicanus%5BTitle%5D)+AND+%22Molecular+Ecology%22%5BJournal%5D)">The role of gene flow in rapid and repeated evolution of cave-related traits in Mexican tetra, Astyanax mexicanus.</a> Molecular Ecology. 27, 4397-4416 (2018).</li>
<li>Rohner, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cryptic+variation+in+morphological+evolution%3A+HSP90+as+a+capacitor+for+loss+of+eyes+in+cavefish%5BTitle%5D)+AND+%22Science+%2880%29%22%5BJournal%5D)">Cryptic variation in morphological evolution: HSP90 as a capacitor for loss of eyes in cavefish.</a> Science (80). 342, 1372-1375 (2013).</li>
<li>McGaugh, S. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+cavefish+genome+reveals+candidate+genes+for+eye+loss%5BTitle%5D)+AND+%22Nature+Communications%22%5BJournal%5D)">The cavefish genome reveals candidate genes for eye loss.</a> Nature Communications. 5, (2014).</li>
<li>Lyon, A., Powers, A. K., Gross, J. B., O'Quin, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Two+-+Three+loci+control+scleral+ossicle+formation+via+epistasis+in+the+cavefish+astyanax+mexicanus%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Two - Three loci control scleral ossicle formation via epistasis in the cavefish astyanax mexicanus.</a> PLoS One. 12, (2017).</li>
<li>Atukorala, A. D. S., Franz-Odendaal, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Genetic+linkage+between+altered+tooth+and+eye+development+in+lens-ablated+Astyanax+mexicanus%5BTitle%5D)+AND+%22Developmental+Biology%22%5BJournal%5D)">Genetic linkage between altered tooth and eye development in lens-ablated Astyanax mexicanus.</a> Developmental Biology. , (2018).</li>
<li>Gore, A. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+epigenetic+mechanism+for+cavefish+eye+degeneration%5BTitle%5D)+AND+%22Nature+Ecology+%26+Evolution%22%5BJournal%5D)">An epigenetic mechanism for cavefish eye degeneration.</a> Nature Ecology & Evolution. 2, 1155-1160 (2018).</li>
<li>Sumner-Rooney, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+kingdom+of+the+blind%3A+disentangling+fundamental+drivers+in+the+evolution+of+eye+loss%5BTitle%5D)+AND+%22Integrative+and+Comparative+Biology%22%5BJournal%5D)">The kingdom of the blind: disentangling fundamental drivers in the evolution of eye loss.</a> Integrative and Comparative Biology. , (2018).</li>
<li>Protas, M. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Genetic+analysis+of+cavefish+reveals+molecular+convergence+in+the+evolution+of+albinism%5BTitle%5D)+AND+%22Nature+Genetics%22%5BJournal%5D)">Genetic analysis of cavefish reveals molecular convergence in the evolution of albinism.</a> Nature Genetics. 38, 107-111 (2006).</li>
<li>Protas, M., Conrad, M., Gross, J. B., Tabin, C., Borowsky, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Regressive+Evolution+in+the+Mexican+Cave+Tetra%2C+Astyanax+mexicanus%5BTitle%5D)+AND+%22Current+Biology%22%5BJournal%5D)">Regressive Evolution in the Mexican Cave Tetra, Astyanax mexicanus.</a> Current Biology. 17, 452-454 (2007).</li>
<li>Protas, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Multi-trait+evolution+in+a+cave+fish%2C+Astyanax+mexicanus%5BTitle%5D)+AND+%22Evolution+%26+Development%22%5BJournal%5D)">Multi-trait evolution in a cave fish, Astyanax mexicanus.</a> Evolution & Development. 10, 196-209 (2008).</li>
<li>Stahl, B. A., Gross, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Alterations+in+Mc1r+gene+expression+are+associated+with+regressive+pigmentation+in+Astyanax+cavefish%5BTitle%5D)+AND+%22Development+Genes+and+Evolution%22%5BJournal%5D)">Alterations in Mc1r gene expression are associated with regressive pigmentation in Astyanax cavefish.</a> Development Genes and Evolution. 225, 367-375 (2015).</li>
<li>McCauley, D. W., Hixon, E., Jeffery, W. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Evolution+of+pigment+cell+regression+in+the+cavefish+Astyanax%3A+A+late+step+in+melanogenesis%5BTitle%5D)+AND+%22Evolution+%26+Development%22%5BJournal%5D)">Evolution of pigment cell regression in the cavefish Astyanax: A late step in melanogenesis.</a> Evolution & Development. 6, 209-218 (2004).</li>
<li>Duboué, E. R., Keene, A. C., Borowsky, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Evolutionary+convergence+on+sleep+loss+in+cavefish+populations%5BTitle%5D)+AND+%22Current+Biology%22%5BJournal%5D)">Evolutionary convergence on sleep loss in cavefish populations.</a> Current Biology. 21, 671-676 (2011).</li>
<li>Yoshizawa, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Distinct+genetic+architecture+underlies+the+emergence+of+sleep+loss+and+prey-seeking+behavior+in+the+Mexican+cavefish%5BTitle%5D)+AND+%22BMC+Biology%22%5BJournal%5D)">Distinct genetic architecture underlies the emergence of sleep loss and prey-seeking behavior in the Mexican cavefish.</a> BMC Biology. 13, 15(2015).</li>
<li>Jaggard, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+lateral+line+confers+evolutionarily+derived+sleep+loss+in+the+Mexican+cavefish%5BTitle%5D)+AND+%22Journal+of+Experimental+Biology%22%5BJournal%5D)">The lateral line confers evolutionarily derived sleep loss in the Mexican cavefish.</a> Journal of Experimental Biology. 220, 284-293 (2017).</li>
<li>Jaggard, J. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Hypocretin+underlies+the+evolution+of+sleep+loss+in+the+Mexican+cavefish%5BTitle%5D)+AND+%22Elife%22%5BJournal%5D)">Hypocretin underlies the evolution of sleep loss in the Mexican cavefish.</a> Elife. 7, (2018).</li>
<li>Duboué, E. R., Borowsky, R. L., Keene, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((%CE%B2-adrenergic+signaling+regulates+evolutionarily+derived+sleep+loss+in+the+mexican+cavefish%5BTitle%5D)+AND+%22Brain%2C+Behavior+and+Evolution%22%5BJournal%5D)">β-adrenergic signaling regulates evolutionarily derived sleep loss in the mexican cavefish.</a> Brain, Behavior and Evolution. 80, 233-243 (2012).</li>
<li>Aspiras, A. C., Rohner, N., Martineau, B., Borowsky, R. L., Tabin, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Melanocortin+4+receptor+mutations+contribute+to+the+adaptation+of+cavefish+to+nutrient-poor+conditions%5BTitle%5D)+AND+%22Proceedings+of+the+National+Academy+of+Sciences+of+the+United+States+of+America%22%5BJournal%5D)">Melanocortin 4 receptor mutations contribute to the adaptation of cavefish to nutrient-poor conditions.</a> Proceedings of the National Academy of Sciences of the United States of America. 112, 9668-9673 (2015).</li>
<li>Riddle, M. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Insulin+resistance+in+cavefish+as+an+adaptation+to+a+nutrient-limited+environment%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Insulin resistance in cavefish as an adaptation to a nutrient-limited environment.</a> Nature. 555, 647-651 (2018).</li>
<li>Xiong, S., Krishnan, J., Peuß, R., Rohner, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Early+adipogenesis+contributes+to+excess+fat+accumulation+in+cave+populations+of+Astyanax+mexicanus%5BTitle%5D)+AND+%22Developmental+Biology%22%5BJournal%5D)">Early adipogenesis contributes to excess fat accumulation in cave populations of Astyanax mexicanus.</a> Developmental Biology. , (2018).</li>
<li>Yoshizawa, M., Gorički, Š, Soares, D., Jeffery, W. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Evolution+of+a+behavioral+shift+mediated+by+superficial+neuromasts+helps+cavefish+find+food+in+darkness%5BTitle%5D)+AND+%22Current+Biology%22%5BJournal%5D)">Evolution of a behavioral shift mediated by superficial neuromasts helps cavefish find food in darkness.</a> Current Biology. 20, 1631-1636 (2010).</li>
<li>Franz-Odendaal, T. A., Hall, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Modularity+and+sense+organs+in+the+blind+cavefish%2C+Astyanax+mexicanus%5BTitle%5D)+AND+%22Evolution+and+Development%22%5BJournal%5D)">Modularity and sense organs in the blind cavefish, Astyanax mexicanus.</a> Evolution and Development. 8, 94-100 (2006).</li>
<li>Hinaux, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Sensory+evolution+in+blind+cavefish+is+driven+by+early+embryonic+events+during+gastrulation+and+neurulation%5BTitle%5D)+AND+%22Development%22%5BJournal%5D)">Sensory evolution in blind cavefish is driven by early embryonic events during gastrulation and neurulation.</a> Development. 143, 4521-4532 (2016).</li>
<li>Blin, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Developmental+evolution+and+developmental+plasticity+of+the+olfactory+epithelium+and+olfactory+skills+in+Mexican+cavefish%5BTitle%5D)+AND+%22Developmental+Biology%22%5BJournal%5D)">Developmental evolution and developmental plasticity of the olfactory epithelium and olfactory skills in Mexican cavefish.</a> Developmental Biology. , (2018).</li>
<li>Lloyd, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Evolutionary+shift+towards+lateral+line+dependent+prey+capture+behavior+in+the+blind+Mexican+cavefish%5BTitle%5D)+AND+%22Developmental+Biology%22%5BJournal%5D)">Evolutionary shift towards lateral line dependent prey capture behavior in the blind Mexican cavefish.</a> Developmental Biology. 441, 328-337 (2018).</li>
<li>Powers, A. K., Boggs, T. E., Gross, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Canal+neuromast+position+prefigures+developmental+patterning+of+the+suborbital+bone+series+in+Astyanax+cave-+and+surface-dwelling+fish%5BTitle%5D)+AND+%22Developmental+Biology%22%5BJournal%5D)">Canal neuromast position prefigures developmental patterning of the suborbital bone series in Astyanax cave- and surface-dwelling fish.</a> Developmental Biology. 441, 252-261 (2018).</li>
<li>Rohner, N. "Out of the Dark" Cavefish Are Entering Biomedical Research. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aRohner%2C+N+%22Zebrafish%2C+Medaka%2C+and+Other+Small+Fishes%22">Zebrafish, Medaka, and Other Small Fishes</a>. , 253-268 (2018).</li>
<li>Yoshizawa, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+evolution+of+a+series+of+behavioral+traits+is+associated+with+autism-risk+genes+in+cavefish%5BTitle%5D)+AND+%22BMC+Evolutionary+Biology%22%5BJournal%5D)">The evolution of a series of behavioral traits is associated with autism-risk genes in cavefish.</a> BMC Evolutionary Biology. 18, 89(2018).</li>
<li>Stockdale, W. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Heart+Regeneration+in+the+Mexican+Cavefish%5BTitle%5D)+AND+%22Cell+Reports%22%5BJournal%5D)">Heart Regeneration in the Mexican Cavefish.</a> Cell Reports. 25, 1997-2007 (2018).</li>
<li>Peuß, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Gamete+Collection+and+In+Vitro+Fertilization+of+Astyanax+mexicanus%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Gamete Collection and In Vitro Fertilization of Astyanax mexicanus.</a> Journal of Visualized Experiments. , (2019).</li>
<li>Luc, H., Sears, C., Raczka, A., Gross, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Wholemount+In+Situ+Hybridization+for+Astyanax+Embryos%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Wholemount In Situ Hybridization for Astyanax Embryos.</a> Journal of Visualized Experiments. , (2019).</li>
<li>Riddle, M., Martineau, B., Peavey, M., Tabin, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Raising+the+Mexican+Tetra+Astyanax+mexicanus+for+Analysis+of+Post-larval+Phenotypes+and+Whole-mount+Immunohistochemistry%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Raising the Mexican Tetra Astyanax mexicanus for Analysis of Post-larval Phenotypes and Whole-mount Immunohistochemistry.</a> Journal of Visualized Experiments. , (2018).</li>
<li>Stahl, B. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Manipulation+of+Gene+Function+in+Mexican+Cavefish%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Manipulation of Gene Function in Mexican Cavefish.</a> Journal of Visualized Experiments. , (2019).</li>
<li>Duboué, E. R., Keene, A. C. Investigating the Evolution of Sleep in the Mexican Cavefish. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aDubou%C3%A9%2C+ER+%22Biology+and+Evolution+of+the+Mexican+Cavefish%22">Biology and Evolution of the Mexican Cavefish</a>. , 291-308 (2015).</li>
<li>Frøland Steindal, I. A., Beale, A. D., Yamamoto, Y., Whitmore, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Development+of+the+Astyanax+mexicanus+circadian+clock+and+non-visual+light+responses%5BTitle%5D)+AND+%22Developmental+Biology%22%5BJournal%5D)">Development of the Astyanax mexicanus circadian clock and non-visual light responses.</a> Developmental Biology. , (2018).</li>
<li>Carlson, B. M., Gross, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Characterization+and+comparison+of+activity+profiles+exhibited+by+the+cave+and+surface+morphotypes+of+the+blind+Mexican+tetra%2C+Astyanax+mexicanus%5BTitle%5D)+AND+%22Comparative+Biochemistry+and+Physiology+Part+C%3A+Toxicology+%26+Pharmacology%22%5BJournal%5D)">Characterization and comparison of activity profiles exhibited by the cave and surface morphotypes of the blind Mexican tetra, Astyanax mexicanus.</a> Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology. 208, 114-129 (2018).</li>
<li>Jaggard, J. B., Lloyd, E., Lopatto, A., Duboue, E. R., Keene, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Automated+Measurements+of+Sleep+and+Locomotor+Activity+in+Mexican+Cavefish%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Automated Measurements of Sleep and Locomotor Activity in Mexican Cavefish.</a> Journal of Visualized Experiments. , (2019).</li>
<li>Kowalko, J. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Convergence+in+feeding+posture+occurs+through+different+genetic+loci+in+independently+evolved+cave+populations+of+Astyanax+mexicanus%5BTitle%5D)+AND+%22Proceedings+of+the+National+Academy+of+Sciences+of+the+United+States+of+America%22%5BJournal%5D)">Convergence in feeding posture occurs through different genetic loci in independently evolved cave populations of Astyanax mexicanus.</a> Proceedings of the National Academy of Sciences of the United States of America. 110, 16933-16938 (2013).</li>
<li>Worsham, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Behavioral+Tracking+and+Neuromast+Imaging+of+Mexican+Cavefish%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Behavioral Tracking and Neuromast Imaging of Mexican Cavefish.</a> Journal of Visualized Experiments. , (2019).</li>
</ol>

作者没有什么可透露的。

尽管我们对动物多样性的起源长期感兴趣，但我们对形态、生理学和行为进化背后的遗传变化的了解有限。墨西哥四角形动物 Astyanax mexicanus 已成为一个强大的模型，可以进一步了解性状如何进化。这种鱼有两种形态型，一种是墨西哥和德克萨斯州南部河流中丰富的有眼河栖形态（表层鱼），另一种是在墨西哥东北部阿布拉山脉地区永远黑暗的石灰岩洞穴中繁衍生息的无眼穴居形态（洞穴鱼）1。有 29 个已知的洞穴种群起源于至少两次独立的地表鱼类入侵 2,3。洞穴和河流变形体一直保持着互惠性，并且很容易在实验室中繁殖，使研究人员能够对响应洞穴中独特选择压力而进化的性状进行基因图谱。例如，缺乏光线导致眼睛4、5、6、7、8、9、色素10、11、12、13、14 和睡眠15、16、17、18、19 退化;营养限制选择暴饮暴食20、体重增加21 和脂肪储存22;在黑暗中导航促进了感知环境的能力的提高 23,24,25,26,27,28。越来越多的研究人员正在利用这种鱼来探索进化的遗传基础。由于洞穴鱼的一些适应性特征对人类有害，因此洞穴鱼也对生物医学研究具有吸引力 21,29,30,31。为了提高可重复性并为新研究人员提供平台，该 JoVE 方法集合提供了用于饲养、性状测量和基因活性检查的技术。
作为起点，Peuß 等人展示的体外受精程序。32 具有多种用途，因为它提供了一种获得 1） 可能难以繁殖的洞穴/地表杂交种，2） 用于显微注射的单细胞阶段胚胎，以及 3） 用于分析胚胎表型的阶段匹配样品的方法。 A. mexicanus 通常在夜间产卵。Peuß 等人。描述一种系统来改变明暗循环并获得在正常工作时间准备收集配子的鱼。他们对配子收集的视觉演示特别有帮助，因为这一步需要仔细的技术。他们的文章对于许多下游应用至关重要，包括胚胎发生过程中基因表达模式的比较。
吕克 等人。33 提供了一种强大的原位杂交方案，针对墨西哥 A. mexicanus 胚胎中的基因表达进行可视化进行了优化。他们使用清单（作为补充文件提供）简化了这个多日过程，并共享故障排除指南。作为这种方法的补充，Riddle 等人。34 展示了如何使用全体免疫组织化学检查孵化鱼的基因表达和蛋白质定位。为了在孵化和摄食开始后对形态型进行有意义的比较，有必要控制生长条件。他们展示了如何通过自然产卵繁殖成虫，并在轮虫饮食中以固定密度饲养幼虫，与其他食物来源相比，轮虫饮食在营养成分和成本方面具有相当大的优势。他们的免疫染色方案最晚在孵化后 12 天成功识别大脑、肠道和胰腺中的抗原，从而可以询问器官发育和功能的后期阶段。重要的是，它们提供了已成功用于 墨西哥曲霉的抗体列表。Luc 等人的联合方法。和里德尔 等人。有助于分析基因作的影响。
在本方法集中，Stahl 等人。35 描述了纵墨西哥 A. mexicanus 基因功能的多种方法，包括：吗啉显微注射、CRISPR-Cas9 介导的诱变和 Tol2 介导的转基因。除了展示注射胚胎的皮秒注射和筛选外，它们还为设计吗啉、引导RNA和转基因提供了有用的指南。他们分享的结果验证了这些技术中的每一种。例如，使用剪接阻断吗啉靶向下丘脑分泌素/食欲素 （HCRT） 会降低穴居鱼的运动活动并增加睡眠;使用 CRISPR/Cas9 编辑表层鱼类的 oca2（洞穴鱼白化病的致病位点）产生白化表层鱼;并插入由使用 Tol2 转基因的泛神经元斑马鱼启动子驱动的基因编码钙指示剂，在墨西哥四核动物中实现近乎普遍的神经元表达。他们指出，斑马鱼开发的许多遗传工具可以直接转移到洞穴鱼身上，因为它们的基因组相似。他们的文章是研究人员对 墨西哥曲霉 进行功能研究的重要资源，并补充了专注于测量表型（包括行为）的其他文章。
洞穴鱼显着减少了睡眠并改变了昼夜节律 36,37,38。这为研究遗传学和环境对脊椎动物睡眠变化的影响提供了一个令人兴奋的机会。贾格德等人。39 展示了如何用廉价材料构建一个系统，这些材料可用于记录不同生命阶段和不同背景下的睡眠。此外，他们还描述了如何在这种设置中自动跟踪鱼类，并获得睡眠和运动的定量测量值。他们的技术适用于高通量药物和基因筛查以及传统的基因图谱研究。
除了睡眠减少之外，洞穴鱼还进化出与觅食相关的运动活动的差异，包括进食角度和振动吸引力的改变 16,30,40。了解这些活动如何演变的核心是将它们与发展变化联系起来。沃沙姆等人。41 提出了记录行为并将其与神经肥大变化相关联的方法;对水流做出反应的外部机械感觉结构。他们首先展示了如何创建腔室来记录振动吸引力和睡眠，并详细描述了如何使用免费软件和可定制的脚本量化行为。接下来，他们演示了如何使用荧光线粒体染色来可视化活体成年鱼中的神经肥大，并再次使用免费软件如何以半自动方式量化神经肥大门的数量。振动吸引力和神经肥大数相关23;他们的方法提供了一个分步协议来询问这种关系。Jaggard 和 Worsham 的方法可用于发现其他行为，也可以应用于其他种类的鱼类。这些方法产生定量且一致的结果，这对于发现行为进化的遗传基础至关重要。
测量和纵基因功能的现代技术已将进化研究从比较研究推向机械研究。结合起来，本方法集中的文章可以作为对墨西哥 A. mexicanus 基因功能进行研究的指南。虽然研究人员已经发现了地表和洞穴变形之间在形态、生理学和行为方面的迷人差异，但仍有许多特征有待详细探索，并且可能还有更多有待发现。随着更多工作的完成，这条小鱼将继续对我们对进化的理解产生重大影响。

小鱼，大问题：墨西哥 tetra 研究的现代技术集合

Little Fish, Big Questions: A Collection of Modern Techniques for Mexican Tetra Research

讨论的文章：
Stahl， B. A. et al. 墨西哥洞穴鱼基因功能的纵。可视化实验杂志。（146） （2019 年）。
Peuß， R. et al. 墨西哥 Astyanax 的配子集合和体外受精。可视化实验杂志。（147） （2019 年）。
Worsham， M. et al. 墨西哥洞穴鱼的行为跟踪和神经桅杆成像。可视化实验杂志。（147） （2019 年）。
Jaggard， J.B.， Lloyd， E.， Lopatto， A.， Duboue， E.R.， Keene， A.C. 墨西哥洞穴鱼睡眠和运动活动的自动测量。可视化实验杂志。（145） （2019 年）。
Luc， H.， Sears， C.， Raczka， A.， Gross， JB Wholemount 原位杂交用于 Astyanax 胚胎。可视化实验杂志。（145） （2019 年）。
Riddle， M.， Martineau， B.， Peavey， M.， Tabin， C. 饲养墨西哥 Tetra Astyanax mexicanus 用于分析幼虫后表型和整体免疫组织化学。可视化实验杂志。（142） （2018 年）。

小鱼，大问题：墨西哥 Tetra 研究的现代技术集合

Articles Discussed:
Stahl, B. A. et al. Manipulation of Gene Function in Mexican Cavefish. Journal of Visualized Experiments. (146) (2019).
Peu&#223;, R. ...

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了解墨西哥盲鱼（Astyanax mexicanus）研究的现代技术集合，涵盖基因操作、行为追踪、神经丘成像及整体原位杂交等方法。这些标准化方案为发育生物学、遗传学和行为神经科学研究提供了可重复的技术支持。

小鱼，大问题：墨西哥 Tetra 研究的现代技术集合

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